brain slicing

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Brain Slicing

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Post on 12-Aug-2015

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Page 1: Brain slicing

Brain Slicing

Page 2: Brain slicing

Advantages

• Precise control over experimental conditions

• Mechanical stability (heartbeat, respiration)

• No blood brain barrier• Easy access of cell location and

identification

Page 3: Brain slicing

Disadvantages

• Lack of inputs and outputs of normal brain

• The slicing process can damage the tissue

• Gets “older” faster• Artificial bathing solution is not

precise

Page 4: Brain slicing

Usage• Study of synaptic activity• Voltage clamp• Iontophoresis• Long term potentiation, adaptation and

kindling

Page 5: Brain slicing

Main steps

1. Chill and oxygenate the sucrose-aCSF (artificial Cerebro-Spinal Fluid)

2. Prepare for brain removal3. Brain removal4. Slice preparation5. Clean up of the workbench

Page 6: Brain slicing

Preparation

• Crush dry ice into powder and put in tray

Page 7: Brain slicing

Rinse beaker

• With distilled water and aCSF

Page 8: Brain slicing

Put beaker in ice tray

• Add water, fill beaker with aCSF

Page 9: Brain slicing

Place air-stone bubblers in beakers

Page 10: Brain slicing

Put in net suspension device

Page 11: Brain slicing

Prepare dissection tray

• Rongeur, scissors, scalpel

Page 12: Brain slicing

Prepare dissection tray

• Filter papers, wipes, super glue

Page 13: Brain slicing

Fill cylinder with 3ml isoflurane

Page 14: Brain slicing

Anesthetization container

Page 15: Brain slicing

Anesthetize the rat

Page 16: Brain slicing

Weigh the rat

• Weighing dish

Page 17: Brain slicing

Decapitate mouse head

• And remove the remaining body

Page 18: Brain slicing

Remove excess blood and tissue

• Dip in petri dish with aCSF

Page 19: Brain slicing

Use a scalpel to cut the scalp

• Use spatula to remove the brain

Page 20: Brain slicing

Put super glue on pedestal

• Then drop the brain

Page 21: Brain slicing

Put slicing pedestal on slicer

Page 22: Brain slicing